PMID:
Front Pharmacol. 2020 ;11:647. Epub 2020 May 8. PMID: 32457629
Abstract Title:
Berberine Improves Glucose and Lipid Metabolism in HepG2 Cells Through AMPKα1 Activation.
Abstract:
Aim: This study is designed to investigate whether or not AMP-activated protein kinaseα1 (AMPKα1) is required for natural product berberine (BBR) to improve glucose and lipid metabolism in HepG2 cells.Methods: AMPKα1 knocked-out (KO,) cells were obtained by co-transfection of the CRISPR/Cas9 KO and HDR (homology-directed repair) plasmid into HepG2 cells, as well as subsequent screen with puromycin. The expression levels of target proteins or mRNAs were determined by western blot or real-time RT-PCR, respectively. Cellular AMPK activity, glucose consumption, lactate release, glucose production, and lipid accumulation were determined by kits.Results: The results showed that thegene was successfully KO in HepG2 cells. Incells, the protein expression of AMPKα1 and phosphorylated-AMPKα1 (p-AMPKα1) disappeared, the level of total AMPKα declined to about 45-50% of wild type (
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